PUBLICATION

Generating Conditional Mutations in Zebrafish Using Gene-trap Mutagenesis

Authors
Maddison, L.A., Lu, J., and Chen, W.
ID
ZDB-PUB-110921-3
Date
2011
Source
Methods in cell biology   104: 1-22 (Chapter)
Registered Authors
Chen, Wenbiao
Keywords
gene-trap, mutagenesis, cre recombinase
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Cloning, Molecular/methods*
  • DNA Transposable Elements
  • Genes, Reporter
  • Genetic Vectors
  • Green Fluorescent Proteins/biosynthesis
  • Green Fluorescent Proteins/genetics
  • Molecular Sequence Data
  • Mutagenesis, Insertional/methods*
  • Recombination, Genetic
  • Zebrafish/genetics*
PubMed
21924154 Full text @ Meth. Cell. Biol.
Abstract
While several mutagenesis methods have been successfully applied in zebrafish, these mutations do not allow tissue- or temporal-specific functional analysis. We have developed a strategy that will allow tissue- or temporal-specific disruption of genes in zebrafish. This strategy combines gene-trap mutagenesis and FlEx modules containing target sites for site-specific recombinases. The gene-trap cassette is highly mutagenic in one orientation and nonmutagenic in the opposite orientation, with different fluorescent proteins as indicators of the orientation. The inclusion of the FlEx modules allows two rounds of stable inversion mediated by the Cre and Flp recombinases. This gene-trap cassette can be easily delivered via transposons. Through large-scale community-wide efforts, broad genome coverage can be obtained. This should allow investigation of cell/tissue-specific gene function of a wide range of genes.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping